银鲫IgM的纯化及其单克隆抗体的制备

Aeromonas hydrophila septicemia was a serious infectious disease occurred in China.The majority harmed fish was crucian carp（Carassius auratus gibelio）.It was reported that immune prevention was the effect measure to prevent this disease using A.hydrophila bacterin.So it is necessary to develop monoclonal antibody（McAb） against the IgM from Crucian carp.In this study,the purification of serum immunoglobulin from Crucian carp was carried out by using Goat IgG Agarose.The concentration of purified IgM was 2.5 mg·mL-1.By using hybridoma-monoclonal antibody technology,hybridoma cell line secreted McAb against the IgM from crucian carp had been established and the characteristics of mab8 had been described.In isotyping analysis,the results showed that it belong to IgG1,it was able to recognize the heavy chain of the IgM on western blot,and did not has any cross reaction with the Ig from 19 different fish species including common carp（Cyprinus carpio）,bighead carp（Aristichthys nobilis）,grass carp（Ctenopharhyngodon idella） and bream（Abramis brama） etc.The results demonstrated that McAb was highly specific and sensitive to the Ig of Crucian carp,and had the great potential to be used for analyzing the structure of crucian carp IgM,monitoring the immunity level of cultured Crucian carp and pathogen diagnosis.